2,4- diaminopyrimidine derivatives

ABSTRACT

There are provided compounds of formula I 
     
       
         
         
             
             
         
       
     
     wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8  and R 9  are as indicated in claim  1 , useful in disorders where ZAP-70 and/or Syk inhibition plays a role or caused by a malfunction of signal cascades connected with FAK.

The present invention relates to pyrimidine derivatives, to processesfor their production, their use as pharmaceuticals and to pharmaceuticalcompositions comprising them.

More particularly the present invention provides in a first aspect, acompound of formula I

wherein

-   X is ═CR⁰— or ═N—;-   each of R⁰, R¹, R², R³ and R⁴ independently is hydrogen; hydroxy;    C₁-C₈alkyl; C₂-C₈alkenyl; C₃-C₈cycloalkyl;    C₃-C₈cycloalkyl-C₁-C₈alkyl; hydroxyC₁-C₈alkyl;    C₁-C₈alkoxyC₁-C₈alkyl; hydroxyC₁-C₈alkoxyC₁-C₈alkyl; arylC₁-C₈alkyl    which optionally may be substituted on the ring by hydroxy,    C₁-C₈alkoxy, carboxy or C₁-C₈alkoxycarbonyl;-   or R³ and R⁴ form together with the nitrogen and carbon atoms to    which they are attached a 5 to 10 membered heterocyclic ring and    comprising additionally 1, 2 or 3 heteroatoms selected from N, O and    S;-   or each of R¹, R² and R³, independently, is halogen;    halo-C₁-C₈alkyl; C₁-C₈alkoxy; halo-C₁-C₈alkoxy; hydroxyC₁-C₈alkoxy;    C₁-C₈alkoxyC₁-C₈alkoxy; aryl; arylC₁-C₈alkoxy; heteroaryl;    heteroaryl-C₁-C₄alkyl; 5 to 10 membered heterocyclic ring; nitro;    carboxy; C₂-C₈alkoxycarbonyl; C₂-C₈alkylcarbonyl;    —N(C₁-C₈alkyl)C(O)C₁-C₈alkyl; —N(R¹⁰)R¹¹; —CON(R¹⁰)R¹¹;    —SO₂N(R¹⁰)R¹¹; or —C₁-C₄-alkylene-SO₂N(R¹⁰)R¹¹; wherein each of R¹⁰    and R¹¹ independently is hydrogen; hydroxy; C₁-C₈alkyl;    C₂-C₈alkenyl; C₃-C₈cycloalkyl; C₃-C₈cycloalkyl-C₁-C₈alkyl;    C₁-C₈alkoxyC₁-C₈alkyl; hydroxyC₁-C₈alkoxyC₁-C₈alkyl;    hydroxyC₁-C₈alkyl; (C₁-C₈alkyl)-carbonyl; arylC₁-C₈alkyl which    optionally may be substituted on the ring by hydroxy, C₁-C₈alkoxy,    carboxy or C₂-C₈alkoxycarbonyl; or 5 to 10 membered heterocyclic    ring;-   or R¹ and R² form together with the C-atoms to which they are    attached aryl or a 5 to 10 membered heteroaryl residue comprising    one or two heteroatoms selected from N, O and S; or-   each of R⁵ and R⁶ independently is hydrogen; halogen; cyano;    C₁-C₈alkyl; halo-C₁-C₈alkyl; C₂-C₈alkenyl; C₂-C₈alkynyl;    C₃-C₈cycloalkyl; C₃-C₈cycloalkylC₁-C₈alkyl; C₈-C₁₀arylC₁-C₈alkyl;-   each of R⁷, R⁸ and R⁹ is independently hydrogen; hydroxy;    C₁-C₈alkyl; C₂-C₈alkenyl; halo-C₁-C₈alkyl; C₁-C₈alkoxy;    C₃-C₈cycloalkyl; C₃-C₈cycloalkylC₁-C₈alkyl; arylC₁-C₈alkyl; —Y—R¹²    wherein Y is a direct bond or O and R¹² is a substituted or    unsubstituted 5, 6 or 7 membered heterocyclic ring comprising 1, 2    or 3 heteroatoms selected from N, O and S; carboxy;    (C₁-C₈alkoxy)-carbonyl; —N(C₁₋₈alkyl)-CO—NR¹⁰R¹¹; —CONR¹⁰R¹¹;    —N(R¹⁰)(R¹¹); —SO₂N(R¹⁰)R¹¹; or R⁷ and R⁸ or R⁸ and R⁹, respectively    form together with the carbon atoms to which they are attached, a 5    or 6 membered heteroaryl comprising 1, 2 or 3 heteroatoms selected    from N, O and S; or a 5 or 6 membered carbocyclic ring.    in free form or salt form.

Any aryl may be phenyl, naphthyl or 1,2,3,4-tetrahydronaphthyl,preferably phenyl. Heteroaryl is an aromatic heterocyclic ring, e.g. a 5or 6 membered aromatic heterocyclic ring, optionally condensed to 1 or 2benzene rings and/or to a further heterocylic ring.

Any heterocyclic ring may be saturated or unsaturated and optionallycondensed to 1 or 2 benzene rings and/or to a further heterocyclic ring.

Examples of heterocyclic rings or heteroaryl include e.g. morpholinyl,piperazinyl, piperidyl, pyrrolidinyl, pyridyl, purinyl, pyrimidinyl,N-methyl-aza-cycloheptan-4-yl, indolyl, quinolinyl, isoquinolinyl,1,2,3,4-tetrahydroquinolinyl, benzothiazolyl, thiazolyl, imidazolyl,benzimidazolyl, benzoxadiazolyl, benzotriazolyl, indanyl, oxadiazolyl,pyrazolyl, triazolyl, and tetrazolyl. Preferred heterocyclic rings orheteroaryl are morpholinyl, piperazinyl, piperidyl, pyrrolidinyl,pyridyl, N-methyl-aza-cycloheptan-4-yl, thiazolyl, imidazolyl andtetrazolyl.

When R⁷ and R⁸ or R⁸ and R⁹ form together with the carbon atoms to whichthey are attached a 5 or 6 membered carbocyclic ring, this maypreferably be cyclopentyl or cyclohexyl.

Halo-alkyl is alkyl wherein one or more H are replaced by halogen, e.g.CF₃.

Any alkyl or alkyl moiety may be linear or branched. C₁₋₈alkyl ispreferably C₁₋₄alkyl. C₁₋₈alkoxy is preferably C₁₋₄alkoxy. Any alkyl,alkoxy, alkenyl, cycloalkyl, heterocyclic ring, aryl or heteroaryl maybe, unless otherwise stated, unsubstituted or substituted by one or moresubstituents selected from halogen; OH; C₁-C₈alkyl; C₁-C₈alkoxy; nitro;cyano; COOH; carbamoyl; C(NH₂)═NOH; —N(R¹⁰)R¹¹; C₃-C₆cycloalkyl; 3 to 7membered heterocyclic ring; phenyl; phenyl-C₁₋₄alkyl; 5 or 6 memberedheteroaryl. When alkyl, alkoxy or alkenyl is substituted, thesubstituent is preferably on the terminal C atom. When the heterocyclicring or heteroaryl is substituted, e.g. as disclosed above, this may beon one or more ring carbon atoms and/or ring nitrogen atom when present.Examples of a substituent on a ring nitrogen atom are e.g. C₁₋₈alkyl,carbamoyl, —C(NH₂)═NOH, —NR¹⁰R¹¹, C₃₋₆cycloalkyl or phenyl-C₁₋₄alkyl,preferably C₁₋₈alkyl, C₃₋₆cycloalkyl or phenyl-C₁₋₄alkyl.

Preferably substituted alkyl or alkoxy as R₇ is alkyl or alkoxysubstituted on the terminal C atom by OH, C₁₋₄alkoxy or a heterocyclicring. When R¹⁰ or R¹¹ is a 5 to 10 membered heterocyclic ring, it may bee.g. thiazolyl.

Halogen may be F, Cl, Br, or I.

Preferably at most one of R¹, R² or R³ is CONR¹⁰R¹¹ or SO₂NR¹⁰R¹¹, morepreferably SO₂NR¹⁰R¹¹.

The compounds of the invention may exist in free form or in salt form,e.g. addition salts with e.g. organic or inorganic acids, for exampletrifluoroacetic acid or hydrochloride acid, or salts obtainable whenthey comprise a carboxy group, e.g. with a base, for example alkalisalts such as sodium, potassium, or substituted or unsubstitutedammonium salts.

In formula I the following significances are preferred independently,collectively or in any combination or sub-combination:

-   (a) X is ═CR⁰;-   (b) R⁰ is hydrogen; halogen, e.g. Cl; C₁-C₄alkyl, e.g. methyl or    ethyl; C₁₋₄alkoxy, e.g. methoxy; preferably hydrogen;-   (c) R¹ is hydrogen; halogen, e.g. Cl or F; OH; C₁-C₈alkyl, e.g.    methyl or ethyl; substituted C₁₋₈alkyl, e.g. terminally OH    substituted C₁₋₈alkyl; —SO₂N(R¹⁰)R¹¹; —N(C₁₋₄alkyl)C(O)C₁₋₄alkyl; a    5 or 6 membered heterocyclic ring optionally substituted on a ring N    atom (when possible); C₁-C₈alkoxy, e.g. methoxy; aryl, e.g. phenyl;    or form together with R² and the C-atoms to which R¹ and R² are    attached 5 to 10 membered aryl or heteroaryl, the latter comprising    1 or 2 nitrogen atoms;-   (d) R² is hydrogen; hydroxy; C₁-C₈alkyl, e.g. methyl or ethyl;    substituted C₁₋₈alkyl, e.g. terminally OH— or C₁₋₄-alkoxy    substituted C₁₋₈alkyl; C₁₋₈alkoxy; C₁-C₄alkoxyC₁-C₈alkoxy;    —CON(R¹⁰)R¹¹, —SO₂N(R¹⁰)R¹¹; or forms together with R¹ and the    C-atoms to which R¹ and R² are attached a 5 to 10 membered aryl or    heteroaryl, the latter comprising 1 or 2 nitrogen atoms;-   (e) R³ is hydrogen; halogen, e.g. Cl, Br; hydroxy; C₁-C₈alkyl, e.g.    methyl or ethyl; substituted C₁₋₈alkyl, e.g. terminally OH    substituted C₁₋₈alkyl; carboxy; CONR¹⁰R¹¹; —SO₂N(R¹⁰)R¹¹; a 5 or 6    membered heterocyclic ring optionally substituted on a ring nitrogen    atom (when possible); or forms together with R⁴ and the N and C    atoms to which R³ and R⁴ are attached a 6 membered heterocyclic    ring;-   (f) R⁴ is hydrogen; or forms together with R³ and the N and C atoms    to which R³ and R⁴ are attached a 6 membered heterocyclic ring;    preferably hydrogen;-   (g) R⁵ is hydrogen; halogen; C₁₋₄alkyl; or CF₃;-   (h) R⁶ is hydrogen;-   (i) R⁷ is hydrogen; hydroxy; C₁₋₄alkyl; substituted C₁₋₄alkyl, e.g.    terminally OH substituted C₁₋₄alkyl; C₁₋₈alkoxy; substituted    C₁₋₈alkoxy, e.g. terminally substituted by OH, C₁₋₄alkoxy or a    heterocyclic ring; NR¹⁰R¹¹; —SO₂N(R¹⁰)R¹¹; —Y—R¹²; CF₃; or R⁷ forms    together with R⁸ and the C-atoms to which R⁷ and R⁸ are attached a 5    membered heteroaryl residue, e.g. bridged by —NH—CH═CH—, —CH═CH—NH—,    —NH—N═CH—, —CH═N—NH—, —NH—N═N— or —N═N—NH—;-   (k) R⁸ is hydrogen; hydroxy; C₁₋₄alkoxy; carboxy; a 5 or 6 membered    heterocyclic ring optionally substituted on a ring C or N atom;    N(C₁₋₄alkyl)-CO—NR¹⁰R¹¹; or forms with R⁷ or R⁹ and the C-atoms to    which R⁷ and R⁸ or Wand R⁹, respectively, are attached a 5 membered    heteroaryl residue, e.g. bridged by —NH—CH═CH—, —CH═CH—NH—,    —NH—N═CH—, —CH═N—NH—, —NH—N═N— or —N═N—NH—;-   (l) R⁹ is hydrogen; C₁₋₄alkoxy; NR¹⁰R¹¹; or forms with R⁸ and the C    atoms to which R⁸ and R⁹ are attached a 5 membered heteroaryl, e.g.    bridged by —NH—CH═CH—, —CH═CH—NH—, —NH—N═CH—, —CH═N—NH—, —NH—N═N— or    —N═N—NH—;-   (m) one of R¹⁰ and R¹¹, independently, is hydrogen or C₁₋₄alkyl and    the other is hydrogen; OH; C₁₋₈alkyl, substituted C₁₋₈alkyl, e.g.    terminally substituted by OH, C₃₋₆cycloalkyl or a heterocyclic ring;    C₂₋₈alkenyl; C₃₋₈cycloalkyl; hydroxyC₁₋₈alkoxyC₁₋₈alkyl; or a 5    membered heterocyclic ring.-   R³ is preferably SO₂NR¹⁰R¹¹.

The present invention also provides a process for the production of acompound of formula I, comprising reacting a compound of formula II

wherein R¹, R², R³, R⁴, R⁵, R⁶ and X are as defined above, and Y is aleaving group, preferably halogen such as bromide, iodine, or inparticular chloride;with a compound of formula III

wherein R⁷, R⁸ and R⁹ are as defined above;and recovering the resulting compound of formula I in free or in form ofa salt, and, where required, converting the compound of formula Iobtained in free form into the desired salt form, or vice versa.

The process may be performed according to methods known in the art, e.g.as described in examples 1 to 4.

The compound of formula II used as starting materials may be obtained byreacting a compound of formula IV

with a compound of formula V

wherein R¹, R², R³, R⁴, R⁵, R⁶, Y and X are as defined above.

The compounds of formula IV and V are known or may be produced inaccordance with known procedures.

The following examples illustrate the invention without any limitation.

The following abbreviations are employed: APC=allophycocyanine,BINAP=2,2′-bis(diphenylphosphino)-1,1′-binaphthyl, cDNA=complementaryDNA, DCM=dichloromethane, DIAD=diisopropyl azodicarboxylate,DMAP=4-dimethylaminopyridine, DMF=dimethylformamide,DMSO=dimethylsulfoxide, DMF=dimethylformamide;Pmc=2,2,5,7,8-pentamethylchroman; tBu=tert.-butyl;DIPCDI=N,N′-diisopropylcarbodiimid; DTT=1,4-dithio-D,L-treitol,DNA=deoxyribonucleic acid, EDTA=ethylenediaminetetra-acetic acid,Lck=lymphoid T-cell protein tyrosine kinase, LAT-11=linker foractivation of T cell, RT=room temperature; RT-PCR=reverse transcriptionpolymerase chain reaction, MS=molecular ion (e.g. M+H¹⁺) determined byelectrospray mass spectroscopy; Eu=europium; ZAP-70=zetachain-associated protein of 70 kD; Syk=p72syk protein tyrosine kinase;SA=streptavidin.

EXAMPLE 12-[2-(1H-Indazol-6-ylamino)-pyrimidin-4-ylamino]-benzenesulfonamide

(a) 2-(2-Chloro-pyrimidin-4-ylamino)-benzenesulfonamide

To a suspension of 8.52 g (49.47 mmol) 2-aminobenzenesulfonamide in 200ml isopropanol is added 22.1 g (148.42 mmol, 3 equivalent)2,4-dichloropyrimidine and 20 ml 10 M hydrochloric acid (200 mmol, 4equivalent). The suspension is stirred at 60° C. for 2 h 15 min. Thereaction mixture is diluted with 2 l ethyl acetate and 500 ml water isadded. The pH is adjusted to 8-9 by addition of sodium bicarbonate. Thelayers are separated and the aqueous layer is reextracted with 500 mlethyl acetate. The organic layers are dried with sodium sulfate,filtered and evaporated to a volume of 300 ml. A crystalline precipitateis formed and removed by filtration (side product). The filtrate isevaporated to 100 ml whereupon the product crystallizes to give2-(2-chloro-pyrimidin-4-ylamino)-benzenesulfonamide (97% purity byHPLC). The mother liquor of this cristallisation is further purified bycolumn chromatography and crystallisation to give further2-(2-chloro-pyrimidin-4-ylamino)-benzenesulfonamide.

(b) 2-[2-(1H-Indazol-6-ylamino)-pyrimidin-4-ylamino]-benzenesulfonamide

To a suspension of 7.25 g (25.46 mmol)2-(2-Chloro-pyrimidin-4-ylamino)-benzenesulfonamide and 4.07 g (30.55mmol, 1.2 equivalent) 6-aminoindazole in 400 ml isopropanol is added 13ml conc. HCl* (130 mmol, 5 equivalent). The suspension is refluxed for 4h 30 min. The reaction mixture is diluted with 1.5 l ethyl acetate and11 water is added. The pH is adjusted to 8-9 by addition of sodiumbicarbonate. The layers are separated and the aqueous layer isre-extracted with 500 ml ethyl acetate. The organic layers are driedwith sodium sulfate, filtered and evaporated to a volume of 300 ml. Acrystalline precipitate (1.01 g) is formed and removed by filtration(side product). The filtrate is purified by chromatography on 200 gsilica gel eluting with ethyl acetate/methanol 95/5 v/v. Uponevaporation crystals are formed which are filtered to give the titlecompound.

¹H NMR (400 MHz, DMSO-d₆): δ 9.42 (s, 1H), 8.34 (d, 1h), 8.28 (d, 1H),8.27 (s, 1H), 7.93 (s, 1H, 7.88 (d, 1H), 7.62 (m, 2H), 7.32 (d, 1H),7.24 (t, 1H), 6.40 (d, 1H).

MS m/z (%): 382 (M+H, 100);

EXAMPLE 22-[2-(3,4,5-Trimethoxy-phenylamino)-pyrimidin-4-ylamino]-benzenesulfonamide

The title compound is prepared from2-(2-chloro-pyrimidin-4-ylamino)-benzenesulfonamide as described inExample 1 using 3,4,5-Trimethoxy-phenylamine instead of 6-aminoindazolein step (b).

¹H NMR (400 MHz, DMSO-d₆): δ 9.18 (s, 1H), 8.22 (d, 1H), 8.17 (d, 1H),7.89 (d, 1H), 7.55 (t, 1H), 7.25 (t, 1H), 7.14 (s, 2H), 6.40 (d, 1H),3.69 (s, 6H), 3.62 (s, 3H). MS m/z (%): 432 (M+H, 100);

EXAMPLE 32-methyl-6-[2-(3,4,5-Trimethoxy-phenylamino)-pyrimidin-4-ylamino]-benzenesulfonamide

The title compound is prepared as described in Example 1 with thedifference that in step (a) 2-amino-6-methyl-benzenesulfonamide is usedinstead of 2-aminobenzenesulfonamide.2-Amino-6-methyl-benzenesulfonamide may be prepared as described byGirard, Y et al.; J. J. Chem. Soc. Perkin Trans. I 1979, 4, 1043-1047:Under an atmosphere of nitrogen m-toluidin (32.1 g, 32.5 ml, 0.30 mmol)is added dropwise to a solution of chlorosulfonyl isocyanate (51.3 ml,83.6 g, 0.59 mmol) in nitroethane (400 ml) at −55-49° C. The cold bathis removed and the mixture allowed to warm to −8° C., whereuponaluminium chloride (51 g, 0.38 mmol) is added. Heating the mixture to100° C. for 20 min forms a clear brown solution, which is cooled to RTand poured on ice. After filtration, washing with ice water and diethylether the precipitate is collected and dissolved in dioxane (300 ml).Water (1000 ml) and conc. HCl (1500 ml) are added to form a suspension,which is heated to 120° C. for 18 h. After cooling to RT the clear brownsolution is washed with diethyl ether/hexane (1400 ml, 1/1 v/v) andadjusted to pH=8 by addition of sodium carbonate. Extraction using ethylacetate (2×1000 ml), washing of the organic phase with water (500 ml)and brine (500 ml), drying (magnesium sulfate) and concentration yieldsa brown solid, which is purified by chromatography on silica usingmethylene chloride/ethanol (100/1 v/v) to yield the desired product as awhite solid.

Melting point: 72-75° C. (Propan-2-ol);

¹H NMR (400 MHz, DMSO-d₆): δ 2.64 (s, 3H, Me), 3.63 (s, 3H, OMe), 3.68(s, 6H, OMe), 6.31 (d, J=5 Hz, 1H, pyrimidine CH), 7.07 (d, J=8 Hz, 1H,arom. CH), 7.15 (s, 2H, arom. CH), 7.40 (t, J=8 Hz, 1H, arom. CH), 7.65(s, 2H, SO₂NH₂), 8.04 (d, J=8 Hz, 1H, arom. CH), 8.12 (d, J=5 Hz, 1H,pyrimidine CH), 9.14 (s, 1H, NH), 9.40 (s, 1H, NH).

MS (ES⁺)m/z: 446 (MH⁺), 468 (MNa⁺)

MS (ES⁻): 444 (M−H)⁻

EXAMPLE 42-Methoxy-6-[2-(3,4,5-trimethoxy-phenylamino)-pyrimidin-4-ylamino]-benzenesulfonamide

The title compound is prepared as described in Example 1 with thedifference that in step (a) 2-amino-6-methoxy-benzenesulfonamide is usedinstead of 2-Amino-6-methyl-benzenesulfonamide.

2-Amino-6-methoxy-benzenesulfonamide may be prepared from 12.3 g ofmeta-anisidine following an analogous procedure as described in Example1a. NMR (400 MHz, DMSO-d₆): δ 3.62 (s, 3H, OMe), 3.69 (s, 6H, OMe), 3.91(s, 3H, OMe), 6.31 (d, J=5 Hz, 1H, pyrimidine CH), 6.86 (d, J=8 Hz, 1H,arom. CH), 7.12 (s, 2H, arom. CH), 7.43 (t, J=8 Hz, 1H, arom. CH), 8.01(d, J=8 Hz, 1H, arom. CH), 8.11 (d, J=5 Hz, 1H, pyrimidine CH), 9.18 (s,1H, NH), 9.79 (br, 1H, NH).

MS (ES⁺): 462.2 (MH⁺), 484.2 (MNa⁺)

MS (ES⁻): 460.3 (M−H)⁻

The compounds of formula X₁

wherein R³, R⁷ and R⁸ are as defined in Table 1, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 1 MS Data Example R³ R⁷ R⁸ *ES+ *ES− *EI  5 —OH—O-(1-methyl)-azacyclohept-4-yl —H 406 404  6 —SO₂NH₂—O-(1-methyl)-azacyclohept-4-yl —H 469.3  7 —SO₂NH₂—O-2-(1-methyl-azacyclopent-2- —H 469.3 yl)-ethyl 8 —OH—O-2-(1-piperidyl)-ethyl —OCH₃ 436.3 434.4 9 —OH—O-2-(1-methyl-azacyclopent-2- —H 406 404 yl)-ethyl 10 —SO₂NH₂—O—CH₂CH₂CH₂-1-imidazolyl —OCH₃ 496 494 11 —SO₂NH₂—O-2-(1-piperidyl)-ethyl —OCH₃ 499.2 497.3 12 —SO₂NH₂—O—CH₂CH₂-1-methyl-imidazol-1- —H 466 464 yl 13 —OH—O-2-[1-(1,2,4-triazolyl)]-ethyl —H 390 388 14 —OH —O-2-hydroxyethyl—OCH₃ 369.4 367.3 15 —SO₂NH₂ —O-2-hydroxyethyl —OCH₃ 431 16 —SO₂NH₂—O-CH₂CH₂-1-imidazolyl —OCH₃ 17 —SO₂NH₂ —O-2-[1-(1,2,4-triazolyl)]-ethyl—H 452 18 —SO₂NH₂ —NH—N═N— 381 19 —SO₂NHCH₃ —O—CH₂CH₂-1-imidazolyl —OCH₃496 494 20 —SO₂NH₂ —O-2-(1-piperidyl)-ethyl —H 469 467 21 —SO₂NH₂—O—CH₂CH₂-1-imidazolyl —H 452 450 22 —OH —O-2-(1-piperidyl)-ethyl —H 40623 —COOH -4-morpholino —H 24 —OH —O—CH₂CH₂CH₂-1-imidazolyl —OCH₃ 433 43125 —SO₂NHCH₃ —CH═N—NH— 396 394 26 —SO₂NH₂ —O-2-(4-morpholino)ethyl —H471 469 27 —SO₂NH₂ —OCH₃ —OCH₃ 402 400 28 —OH —O-2-(4-morpholino)ethyl—H 408 406 29 —SO₂NH₂ —CH═N—NH— 381 30 —SO₂NHCH₃ —O—CH₂CH₂-1-imidazolyl—H 31 —COOH amino —H 322 32 —SO₂NH₂ —O—CH₂CH₂CH₂-1-imidazolyl —H 466.2464.3 33 —COOH —N(CH₃)₂ —H 34 -5-(1,2,3,4-tetrazolyl) —NH—C(O)CH₃ —H 388386 35 —SO₂NHCH₃ —NH—N═N— 36 —COOH —OH —H 37 —COOH —H -4-piperidyl 38—COOH —CH₂—OH —H 39 —OH —O—CH₂CH₂-1-imidazolyl —OCH3 40 —SO₂NH—CH₂CH₂—OH—O—CH₂CH₂-1-imidazolyl —H 496 494 41 —C(O)NH₂ amino —H 321 42 —SO₂NH₂—CH═CH—NH— 381 43 -5-(1,2,3,4-tetrazolyl) —NHCH₂-3-pyridyl —H 435 44—SO₂NH₂ —NH—CH═CH— 379 45 —COOH —H -4- morpholino 46 —COOH —H -1-(4-amino)- piperidyl 47 —SO₂NH₂ —OCH₃ —H 372 370 48 —SO₂N(CH₃)₂—O—CH₂CH₂-1-imidazolyl —H 480 478

The compounds of formula X₂

wherein R³ and R⁸ are as defined in Table 2, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 2 MS Data Example R³ R⁸ *ES+ *ES− 49 —COOH —OCH₃ 397 395 50—SO₂NH₂ —OH 51 —SO₂NHCH₃ —OCH₃ 52 -5-(1,2,3,4-tetrazolyl) —OCH₃ 421 53—SO₂NH-cyclopropyl —OCH₃ 472.2 470.3 54 —C(O)NHOH —OCH₃ 412 410 55—SO₂NH—CH₂CH₂—OH —OCH₃ 476 474 56 —SO₂N(CH₃)₂ —OCH₃ 460.3 458.3 57 —OH—OCH₃ 369 367 58 —SO₂NH—CH₂CH₂CH₃ —OCH₃ 474 472 59 —CH₂OH —OCH₃ 60—SO₂NH₂ —H 402

The compounds of formula X₃

wherein R¹, R⁷, R⁸ and R⁹ are as defined in Table 3, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 3 MS Data Example R¹ R⁷ R⁸ R⁹ *ES+ *ES− 61—SO₂NH—CH₂CH₂—O—CH₂CH₂—OH —H —N(CH₃)—C(O)CH₃ —H 62 —SO₂NH₂ —OCH₃ —OCH₃—OCH₃ 63 —SO₂NH₂ —O—CH₂CH₂-1-imidazolyl —OCH₃ —H 64—SO₂NH—CH₂CH₂—O—CH₂CH₂—OH —OCH₃ —OCH₃ —OCH₃ 520 518 65 —N(CH₃) C(O)CH₃—OCH₃ —OCH₃ —OCH₃ 424 422 66 —CH₂CH₂—OH —SO₂NH—CH₂CH₂CH₂CH₃ —H —H 67—SO₂NH₂ —OCH₃ —H —OCH₃ 68 —SO₂NH₂ —O—CH₂CH₂-1-imidazolyl —H —H 69—CH₂CH₂—OH —O—CH₂CH₂-1-imidazolyl —H —H 70 —CH₂CH₂—OH —OCH₃ —H —OCH₃ 71—SO₂NH₂ —OH —H —H 72 —O—CH₂CH₂—OH —O—CH₂CH₂-1-imidazolyl —H —H 73—SO₂NH-2-thiazolyl —OCH₃ —OCH₃ —OCH₃ 515 513

The compounds of formula X₄

wherein R², R⁵, R⁷, R⁸ and R⁹ are as defined in Table 4, may be preparedby following the procedure of Example 1 but using the appropriatestarting materials.

TABLE 4 MS Data Example R² R⁵ R⁷ R⁸ R⁹ *ES+ *ES− 74 —SO₂NH-2-propenyl —H—OCH₃ —OCH₃ —OCH₃ 472 470 75 —SO₂NH₂ —H —OCH₃ —OCH₃ —OCH₃ 76 —OH —H—O-(1-methyl)- —H —H 406.3 404.3 azacyclohept-4-yl 77 —OH —H—O—CH₂CH₂—OH —OCH₃ —H 369 367 78 —SO₂NH₂ —Br —OCH₃ —OCH₃ —OCH₃ 510.1/508.1/ 512.1 510.2 79 —SO₂NH₂ —H —CH═N—NH— —H 382 80 —SO₂NH₂ —CH₃ —OCH₃—OCH₃ —OCH₃ 446 444 81 —SO₂NH₂ —H —O—CH₂CH₂-1-imidazolyl —OCH₃ —H 482480 82 —OH —H —O—CH₂CH₂-1-piperidyl —OCH₃ —H 436.3 434.3 83 —OH —H—O—CH₂CH₂-1-imidazolyl —OCH₃ —H 419 417 84 —SO₂NH₂ —H—O—CH₂CH₂-1-imidazolyl —H —H 452 450 85 —CH₃ —C≡N —OCH₃ —OCH₃ —OCH₃ 39286 —SO₂NH₂ —H —NH—N═CH— —H 382 87 —OH —H —OCH₃ —OCH₃ —OCH₃ 369 367 88—SO₂NHCH₃ —CH₃ —OCH₃ —OCH₃ —OCH₃ 460 458 89 —OH —H —OH —COOH —OCH₃ 90—OH —H —O—CH₂CH₂-1-piperidyl —H —H 406 404 91 —SO₂NH-2-propenyl —H—O—CH₂CH₂-1-imidazolyl —H —H 492.3 490.3 92 —SO₂NH₂ —Br—O—CH₂CH₂-1-(1-methyl)- —H —H 544.1/ 542/ imidazolyl 546 544.2 93—SO₂NH₂ —H —O—CH₂CH₂—OH —OCH₃ —H 94 —OH —H —O-(1-methyl)- —H —Hazacyclopent-2-yl 95 —OH —H —O—CH₂CH₂-1-imidazolyl —H —H 389 387 96 —OH—H —O—CH₂CH₂CH₂-1- —OCH₃ —H 433.4 431.4 imidazolyl 97 —SO₂NH₂ —H —OCH₃—H —OCH₃ 98 —OH —H —OCH₃ —OCH₃ —H 339 337 99 —SO₂NHCH₂—CH₂CH₂CH₃ —H—OCH₃ —OCH₃ —OCH₃ 488 486 100 —SO₂NH—CH₃ —CH₃ —O—CH₂CH₂-1-imidazolyl—OCH₃ —H 510 508 101 —SO₂NHCH₂—CH₂CH₂CH₃ —H —O—CH₂CH₂-1-imidazolyl —H —H08 506 102 —OH —H —O—CH₂CH₂-4-morpholino —H —H 408 103 —OH —H —NH—N═CH——H 319 317 104 —OH —H —CHN—NH— —H 319 317 105 —OH —H—O—CH₂CH₂-1-imidazolyl —H —H 106 —SO₂NH—CH₃ —CH₂—CH₃ —OCH₃ —OCH₃ —OCH₃474.3 472.3 107 —SO₂NH₂ —H —OCH₃ —OCH₃ —OCH₃

The compounds of formula X₅

wherein R⁰, R¹, R², R³ and R⁴ are as defined in Table 5, may be preparedby following the procedure of Example 1 but using the appropriatestarting materials.

TABLE 5 MS Data Example R⁰ R¹ R² R³ R⁴ *ES+ *ES− 108 —H —OCH₃ —OH —H —H109 —H nitro —H —OH —H 414 412 110 —H —N═CH—CH═CH— —H —H 111 —H—CH═N—NH— —H —H 393 391 112 —H —NH—N═CH— —H —H 393 113 —H —H —OH—CH₂CH₂CH₂— 409 407 114 —CH₃ —H —CH₃ —OH —H 397 115 —H phenyl —H —SO₂NH₂—H 508 506 116 —CH₃ —H —H —SO₂NH₂ —H 446 444

The compounds of formula X₆

wherein R⁵, R⁷, R⁸ and R⁹ are as defined in Table 6, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 6 Example R⁵ R⁷ R⁸ R⁹ *Es+ *Es− 117 —CH₃ —O—CH₂CH₂-1-imidazolyl —H—H 466 118 —CH₂CH₃ —OCH₃ —OCH₃ —OCH₃ 460 458 119 —Br —NH—N═CH— —H 461120 —CH₃ —O—CH₂CH₂-1-imidazolyl —OCH₃ —H 496 121 —CH₃ —OCH₃ —OCH₃ —OCH₃446 122 —CH₃ —N═N—NH— —H 397.2 395.2 123 —CH₃—O—CH₂CH₂-1-methyl-imidazol-1-yl —H —H 480 124 —Br —CH═N—NH— —H 461.3458.1/ 460 125 —CH₃ —NH—N═CH— —H 396 126 —Br—OCH₂CH₂-(4-methyl-piperazin-1-yl) —H —H 562/ 560/ 564 562

The compounds of formula X₇

wherein R¹, R², R³, R⁷ and R⁸ are as defined in Table 7, may be preparedby following the procedure of Example 1 but using the appropriatestarting materials.

TABLE 7 Ex R¹ R² R³ R⁷ R⁸ *ES+ *ES− 127 —OCH₃ —OH —H —OH —OCH₃ 128 —H—CH₃ —SO₂NH₂ —O—CH₂CH₂-1-imidazolyl —H 466 464 129 —OCH₃ —OH —H—O—CH₂CH₂-1-imidazolyl —OCH₃ 130 —OCH₃ —OH —H —O—CH₂CH₂—OH —OCH₃ 399 397131 —OCH₃ —OH —H —O-(1-methyl-azacyclohept-4-yl) —H 436 132 —CH₃ —H—SO₂NH₂ —O—CH₂CH₂-1-imidazolyl —H 466 464 133 —OCH₃ —OH —H—O—CH₂CH₂-(1-methyl)- —H 436 434 azacyclopent-2-yl 134 —OCH₃ OH —H —CF₃—H 135 —N═CH—CH═CH— —H —O—CH₂CH₂-1-imidazolyl —OCH₃ 136 —OCH₃ —OH —H—O—CH₂CH₂CH₂-1-imidazolyl —OCH₃ 463 461 137 —OCH₃ —OH —H—O—CH₂CH₂-1-piperidyl —OCH₃ 466.4 464.4 138 —CH═N—NH— —H —NH—N═CH— 139—CH═N—NH— —H —CH—N═NH— 140 —OCH₃ —H —H —O—CH₂CH₂-1-piperidyl —H 436 434141 —H —OCH₃ —SO₂NH₂ —O—CH₂CH₂-1-pyrrolidinyl —H 485.3 483.3 142 —H—OCH₃ —SO₂NH₂ —O—CH₂CH₂-1-pyrrolidinyl —CH₃ 499.2 497.3 143 —H —OCH₃—SO₂NH₂ —O—CH₂CH₂CH₂-morpholino —OCH₃ 545.2 545.3 144 —H —OCH(CH₃)₂—SO₂NH₂ —O—CH₂CH₂-(4-methyl-piperazin- —OCH₃ 572.2 570.3 1-yl) 145 —H—OCH₃ —SO₂NH₂ —O—CH₂CH₂-1-piperidinyl —H 499.2 497.3 146 —CH₃ —OCH₃—SO₂NH₂ —O—CH₂CH₂CH₂-1-pyrrolidinyl —OCH₃ 543.2 147 —CH₃ —OCH₃ —SO₂NH₂—O—CH₂CH₂CH₂-1-pyrrolidinyl —H 513.2 511.2 148 —H —OCH(CH₃)₂ —SO₂NH₂—O—CH₂CH₂-1-piperidinyl —H 527.2 525.3 149 —H —CH₃ —SO₂NH₂ —N(CH₃)₂—OCH₃ 429.3 427.3 150 —CH₃ —CH₃ —SO₂NH₂ —O—CH₂CH₂CH₂-1-pyrrolidinyl—OCH₃ 527.2 525.3 151 —OCH₃ —H —SO₂NH₂ —O—CH₂CH₂CH₂-1-pyrrolidinyl —OCH₃529.2 527.3 152 —H —F —SO₂NH₂ —N(CH₃)₂ —OCH₃ 433.1 153 —H —CH₃ —SO₂NH₂—O—CH₂CH₂-(1-methyl-pyrrolidin- —H 2-yl) 154 —H —OCH₃ —SO₂NH₂—O—CH₂CH₂—OH —H 432.2 430.2 155 —H —CH₃ —SO₂NH₂—O—CH₂CH₂-(1-methyl-pyrrolidin- —OCH₃ 513.2 511.3 2-yl) 156 —OCH₃ —H—SO₂NH₂ —O—CH₂CH₂-1-piperidinyl —H 499.2 497.3 157 —OCH₃ —H —SO₂NH₂—O—CH₂CH₂-1-pyrrolidinyl —OCH₃ 515.2 513.2 158 —H —CH₃ —SO₂NH₂—O—CH₂CH₂—OH —OCH₃ 446.2 444.2 159 —OC₂H₅ —H —SO₂NH₂—O—CH₂CH₂-1-pyrrolidinyl —CH₃ 513.3 511.3 160 —OCH₃ —OCH₃ —SO₂NH₂—O—CH₂CH₂-(4-methyl-piperazin- —OCH₃ 574.2 572.2 1-yl) 161 —H —Cl—SO₂NH₂ -(4-methyl-piperazin-1-yl) —H 474.5 472.5 162 —H —CH₃ —SO₂NH₂—O—CH₂CH₂-(4-cyclopentyl- —H 552.3 550.3 piperazin-1-yl) 163—CH═CH—CH═CH— —SO₂NH₂ -(4-methyl-piperazin-1-yl) —H 490.5 488.4 164 —H—H —SO₂NH₂ —O—CH₂CH₂-piperazin-1-yl —H 470.2 468.3 165 —H —OCH₃ —SO₂NH₂—H —OCH₃ 402.2 400.2 166 —H —OCH₃ —SO₂NH₂ —O—CH₂CH₂-(4-benzyl-piperazin-—H 590.3 588.3 1-yl) 167 —CH₃ —H —SO₂NH₂ —O—CH₂CH₂-1-pyrrolidinyl —H469.2 467.3 168 —Br —H —SO₂NH₂ —O—CH₂CH₂-1-piperidinyl —H 549.1 547.2

The compounds of formula X₈

wherein R¹, R², R³ and R⁸ are as defined in Table 8, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 8 Ex R¹ R² R³ R⁸ *ES+ *ES− 169 4-morpholino —H —H —H 170 —CH═N—NH——H —H 363 361 171 —OCH₃ —OH —H —H 172 —CH₃ —H —SO₂NH₂ —OCH₃ 446

The compounds of formula X₉

wherein R⁷, R⁸ and R⁹ are as defined in Table 9, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 9 Example R⁷ R⁸ R⁹ *ES+ *ES− 173 —O—CH₂CH₂-1-piperidyl —OCH₃ —H470.3 468.3 174 —O-(1-methyl-azacyclohept-4-yl) —H —H 440 175—O-(1-methyl-azacyclopent-2-yl) —H —H 440 438 176—O—CH₂CH₂—CH₂-1-imidazolyl —OCH₃ —H 467 465 177 —OCH₃ —OCH₃ —OCH₃ 178—O—CH₂CH₂-1-(1,2,4-triazolyl) —H —H 424 422 179 —O—CH₂CH₂-1-piperidyl —H—H 180 —O—CH₂CH₂—OH —OCH₃ —H 181 —O—CH₂CH₂-4-morpholino —H —H 442 440182 —O—CH₂CH₂CH₂-1-imidazolyl —H —H

The compounds of formula X₁₀

wherein R¹, R⁷ and R⁹ are as defined in Table 10, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 10 EX R¹ R⁷ R⁹ *ES+ *ES− 183 —CH₂CH₂—OH —OCH₃ —OCH₃ 411 409 184—SO₂NH₂ —O—CH₂CH₂-1- —H 496.3 494.3 imidazolyl

The compounds of formula X₁₁

wherein R⁸ is —OCH₃ (Example 185) or —OH (Example 186), may be preparedby following the procedure of Example 1 but using the appropriatestarting materials.

The compounds of formula X₁₂

wherein R⁰, R¹, R⁷, R⁸ and R⁹ are as defined in Table 12, may beprepared by following the procedure of Example 1 but using theappropriate starting materials.

TABLE 12 Example R⁰ R¹ R⁷ R⁸ R⁹ 187 —H —H —H —SO₂NH₂ —H 188 —H —H —H —H—CH₃ 189 —H —H —H —CH₃ —H 190 —H —F —OCH₃ —OCH₃ —OCH₃ 191 —H —H —H —CH₃—CH₃ 192 —H —H —CH₃ —H —CH₃ 193 —H —H —OCH₃ —CH₃ —H 194 —H —H —H —H—N(CH₃)₂ 195 —H —H —OCH(CH₃)₂ —H —H 196 —H —H —H —OCH(CH₃)₂ —H 197 —H —H—CH(CH₃)₂ —H —H 198 —H —H —H —CH═N—NH— 199 —H —H —OCH₃ —CH₃ —OCH₃ 200—OCH₃ —H —OCH₃ —OCH₃ —OCH₃ 201 —H —H —H —H —H 202 —CH₃ —Cl —OCH₃ —OCH₃—OCH₃ 203 —H —H —H —H —CF₃ 204 —Cl —CH₃ —OCH₃ —OCH₃ —OCH₃ 205 —H —H —H—NH—CH═N— 206 —H —H —H —N(—CH₂CH₂CH₂-4- morpholino)-CH═CH— 207 —H —H—CH₂CH₂—CH₂— —H

The compounds of formula X₁₃

wherein R¹, R², R³ and R⁵ are as defined in Table 13, may be prepared byfollowing the procedure of Example 1 but using the appropriate startingmaterials.

TABLE 13 Example R¹ R² R³ R⁵ *ES+ *ES− 208 —H —H —SO₂NHCH₃ —CF₃ 514.0209 —H —H —SO₂NHC₃H₇ —Br 210 —H —H —SO₂NH—CH₂CH— —Br cyclopropyl 211 —H—H —SO₂NHCH₃ —CH₃ 212 —H —H —SO₂N(CH₃)₂ —Br 213 —H —H —SO₂NHCH₃ —Cl 214—H —H —SO₂NHCH₃ —I 215 —H —H —SO₂NHCH₃ —Br 216 —CH₃ —OCH₃ —SO₂NH₂ —H 476474 217 —H piperidino —SO₂NH₂ —H 515.5 513.4 218 —H morpholino —SO₂NH₂—H 517.4 515.4 219 —H —C₂H₅ —SO₂NH₂ —H 220 —H —CH₃ —SO₂NH₂ —Cl 221 —H—CH₃ —SO₂NHCH₃ —H 460.4 222 —H phenyl —SO₂NH₂ —H 508.2 506.3

The compounds of formula X₁₄

wherein R², R³, R⁵, R⁷, R⁸ and R⁹ are as defined in Table 14, may beprepared by following the procedure of Example 1 but using theappropriate starting materials.

TABLE 14 Ex R² R³ R⁵ R⁷ R⁸ R⁹ *ES+ *ES− 223 —OCH₃ —SO₂NH₂ —H —H—CH═N—N(CH₃)— 424 224 —OCH₃ —SO₂NH₂ —H —O—CH₂CH₂—OCH₃ —OCH₃ —H 476.2474.3 225 —OCH(CH₃)₂ —SO₂NH₂ —H —O—CH₂CH₂— —OCH₃ —H 551.2 555.3piperidino 226 —OCH₃ —SO₂NH₂ —H —O—CH₂CH₂-(4- —H —H 514.3 512.3methyl-piperazin-1-yl)- 227 —OCH₃ —SO₂NH₂ —H -morpholino —OCH₃ —H 487.1485.2 228 —CH₃ —SO₂NH₂ —H —O—CH₂CH₂CH₂- —OCH₃ —H 527.3 piperidino 229—CH₃ —SO₂NH₂ —H —O—CH₂CH₂CH₂-1- —OCH₃ —H 513.2 511.3 pyrrolidinyl 230—O—CH₂CH₂—OCH₃ —SO₂NH₂ —H —H —CH═N—N(CH₃)— 539 537 231 -(4-methyl-—SO₂NH₂ —H —OCH₃ —OCH₃ —OCH₃ 530.4 528.4 piperazin-1-yl) 232 —OCH₃—SO₂NH₂ —H —O—CH₂CH₂—OH —OCH₃ —H 462.2 460.3 233 —OCH₃ —SO₂NH₂ —Br—O—CH₂CH₂—OCH₃ —OCH₃ —H 234 —CH₃ —SO₂NH₂ —H —O—CH₂CH₂-(4- —OCH₃ —H 528.2526.3 methyl-piperazin-1-yl) 235 —CH₃ —SO₂NH₂ —H —O—CH₂CH₂—N(CH₃)₂ —H —H443.2 441.3 236 —H —SO₂NH₂ —H —O—CH₂CH₂-1- —OCH₃ —H 485.2 483.3pyrrolidinyl 237 —CH₃ —SO₂NH₂ —H —H —N(CH₃)—N═CH— 410 238 —CH₃ —SO₂NH₂—H —CH₃ —OCH₃ OCH₃ 239 —CH₃ —SO₂NH₂ —Br —O—CH₂CH₂—OCH₃ —OCH₃ —H 538/540240 —OCH₃ —SO₂NH₂ —H —OCH₃ —H —H 402.2 400.2 241 —H —SO₂NH₂ —H —H—CO—NH—CH₂CH₂—OCH₃ —H ES+ means electrospray MS positive mode; ES− meanselectrospray MS negative mode; and EL means electron impact MS.

The compounds of formula I and their pharmaceutically acceptable salts,exhibit valuable pharmacological properties when tested in in vitroassays, and are therefore useful as pharmaceuticals.

In particular the compounds of the invention exhibit ZAP-70 (zetachain-associated protein of 70 kD), Focal Adhesion Kinase (FAK) and/orSyk protein tyrosine kinases inhibiting activity. More particularly thecompounds of the invention are active at the human ZAP-70, FAK and/orSyk protein tyrosine kinases. ZAP-70, FAK and/or Syk protein tyrosinekinase interaction of the compounds of the invention may be demonstratedby their ability to prevent phosphorylation of e.g. LAT-11 (SEQ IDNO: 1) by human ZAP-70 protein tyrosine kinase, to preventphosphorylation of e.g. Biot-Y397 (SEQ ID NO:2) by human FAK proteintyrosine kinase, and/or to prevent phosphorylation of e.g. polymericglutamic acid-tyrosine (Glu, Tyr) by human Syk protein tyrosine kinasein, e.g. aqueous solution, e.g. as demonstrated in accordance with thefollowing test methods.

1. Cell-Free Kinase Assays: ZAP-70 and Syk Kinase Assays

ZAP-70, Lck and Syk are commercially available from UpstateBiotechnology, Lake Placid, N.Y.

Preparation of LAT-11 (SEQ ID NO:1):

The peptide LAT-11 used as a substrate in the ZAP-70 kinase assay may beprepared as disclosed in Example 1A of WO 02/12275, the contents ofwhich, particularly with reference to Example 1A, is incorporated hereinby reference.

ZAP-70 Kinase Assay:

The activities of the agents of invention are determined in a homogenousZAP-70 kinase assay based on time-resolved fluorescence resonance energytransfer. Briefly, 80 nM ZAP-70 are incubated with 80 nM Lck and 4 μMATP in ZAP-70 kinase buffer (20 mM Tris, pH 7.5, 10 μM Na₃VO₄, 1 mM DTT,1 mM MnCl₂, 0.01% bovine serum albumin, 0.05% Tween 20) for 1 hour atroom temperature in a siliconized polypropylene tube. Then, theselective Lck inhibitor PP2(4-amino-5-(4-chloro-phenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine;Alexis Biochemicals) is added (final concentration 1.2 μM) and incubatedfor further 10 min. Ten μl of this solution is mixed with the 10 μlbiotinylated peptide LAT-11 (1 μM) as substrate and 20 μl of serialdilutions of inhibitors and incubated for 4 hours at room temperature.The kinase reaction is terminated with 10 μl of a 10 mM EDTA solution indetection buffer (20 mM Tris, pH 7.5, 0.01% bovine serum albumin, 0.05%Tween 20). The detection phase is performed by addition of 50 μleuropium (Eu)-labelled anti-phosphotyrosine antibody (e.g. Eu-PT66;final concentration 0.125 nM; Advant/Wallac) and 50 μlstreptavidin-allophycocyanine (SA-APC; final concentration 40 nM) indetection buffer. After 1 hour incubation at room temperaturefluorescence is measured, e.g., on the Victor2 Multilabel Counter(Wallac) at 665 nm. Background values (low control) are obtained in theabsence of test samples and ATP and are subtracted from all values.Signals obtained in the absence of test samples are taken as 100% (highcontrol). The inhibition obtained in the presence of test compounds wascalculated as percent inhibition of the high control. The concentrationof test compounds resulting in 50% inhibition (IC₅₀) was determined fromthe dose-response curves. In this assay, the compounds of the inventionhave IC₅₀ values in the range of 10 nM to 2 μM, preferably from 10 nM to100 nM. Compound of Example 4 shows an IC₅₀ value of 12 nM.

Syk Kinase Assay:

The activities of the agents of invention are determined in aheterogenous Syk kinase assay based on the dissociation-enhancedlanthanide fluoroimmunoassay (DELFIA) technology. This method utilizeseuropium chelate-labelled anti-phosphotyrosine antibodies to detectphosphate transfer by Syk to a polymeric glutamic acid-tyrosine (Glu,Tyr) substrate coated onto microtiter plates as described (Braunwalder AF, Yarwood D R, Sills M A, Lipson K E. Measurement of the proteintyrosine kinase activity of c-src using time-resolved fluorometry ofeuropium chelates. Anal. Biochem. 1996; 238(2):159-64). The amount ofphosphorylation is then quantified with time-resolved,dissociation-enhanced fluorescence. Briefly, hundred μl of poly (Glu,Tyr) (4:1; 2 μg/ml in phosphate-buffered saline, PBS) are coated toELISA plates overnight at room temperature. The poly (Glu, Tyr) solutionis removed and 250 μl of 1% bovine serum albumin in PBS are added forone hour at room temperature. Plates are then washed three times with350 μl of washing buffer (25 mM Tris-HCl, pH 7.4 containing 0.03%Tween-20). The kinase reaction is performed for one hour at roomtemperature by mixing serial dilutions of inhibitors in 30 μl with 30 μlof Syk kinase (20 ng/ml) and ATP (1 μM) in kinase buffer (20 mM Tris, pH7.5, 10 μM Na₃VO₄, 1 mM DTT, 10 mM MnCl₂, 2 mM MgCl₂, 0.01% bovine serumalbumin, 0.05% Tween 20). After washing the plates four times asdescribed above 60 μl DELFIA europium N1-labelled anti-phosphotyrosineantibody PY20 (Advant/Wallac) are added (100 ng/ml in 50 mM Tris-HCl,pH7.4, 150 mM NaCl, 20 μM Titriplex V, 0.2% bovine serum albumine, 0.05%Tween-20) and incubated for one hour at room temperature. Plates arewashed eight times and 60 μl enhancement solution (Wallac) are added.Fluorescence is determined at 615 nm (Victor2; Wallac). High controlvalues (100% signal) are obtained in absence of test samples and lowcontrol values (background) in absence of test samples and ATP. Lowcontrols were subtracted from all values. The inhibition obtained in thepresence of test compounds was calculated as percent inhibition of thehigh control. The concentration of test compounds resulting in 50%inhibition (IC₅₀) was determined from the dose-response curves. In thisassay, the compounds of the invention have IC₅₀ values in the range of100 nM to 10 μM, preferably from 100 to 1 μM. Compound of Example 128has an IC₅₀ value of 150 nM.

2. Allogeneic Mixed Lymphocyte Reaction (MLR)

Compounds of the invention exhibit T cell inhibiting activity. Moreparticular the compounds of the invention prevent T cell activationand/or proliferation in e.g. aqueous solution, e.g. as demonstrated inaccordance with the following test method. The two-way MLR is performedaccording to standard procedures (J. Immunol. Methods, 1973, 2, 279 andMeo T. et al., Immunological Methods, New York, Academic Press, 1979,227-39). Briefly, spleen cells from CBA and BALB/c mice (1.6×10⁵ cellsfrom each strain per well in flat bottom tissue culture microtiterplates, 3.2×10⁵ in total) are incubated in RPMI medium containing 10%FCS, 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco BRL, Basel,Switzerland), 50 μM 2-mercaptoethanol (Fluka, Buchs, Switzerland) andserially diluted compounds. Seven three-fold dilution steps induplicates per test compound are performed. After four days ofincubation 1 μCi ³H-thymidine is added. Cells are harvested after anadditional five-hour incubation period, and incorporated ³H-thymidine isdetermined according to standard procedures. Background values (lowcontrol) of the MLR are the proliferation of BALB/c cells alone. Lowcontrols are subtracted from all values. High controls without anysample are taken as 100% proliferation. Percent inhibition by thesamples is calculated, and the concentrations required for 50%inhibition (IC₅₀ values) are determined. In this assay, the compounds ofthe invention have IC₅₀ values in the range of 10 nM to 10 μM,preferably from 10 nM to 100 nM. Compound of Example 120 shows an IC₅₀value of 13 nM.

3. FAK Assay

All steps are performed in a 96-well black microtiter plate. Purifiedrecombinant hexahistidine-tagged human FAK kinase domain is diluted withdilution buffer (50 mM HEPES, pH 7.5, 0.01% BSA, 0.05% Tween-20 inwater) to a concentration of 94 ng/mL (2.5 nM). The reaction mixture isprepared by mixing 10 μL 5× kinase buffer (250 mM HEPES, pH 7.5, 50 μMNa₃VO₄, 5 mM DTT, 10 mM MgCl₂, 50 mM MnCl₂, 0.05% BSA, 0.25% Tween-20 inwater), 20 μL water, 5 μL of 4 μM biotinylated peptide substrate(Biot-Y397) in aqueous solution, 5 μL of test compound in DMSO, and 5 μLof recombinant enzyme solution and incubated for 30 min at roomtemperature. The enzyme reaction is started by addition of 5 μL of 5 μMATP in water and the mixture is incubated for 3 hours at 37° C. Thereaction is terminated by addition of 200 μL of detection mixture (1 nMEu-PT66, 2.5 μg/mL SA-(SL)APC, 6.25 mM EDTA in dilution buffer), and theFRET signal from europium to allophycocyanin is measured by ARVOsx+L(Perkin Elmer) after 30 min of incubation at room temperature. The ratioof fluorescence intensity of 665 nm to 615 nm is used as a FRET signalfor data analysis in order to cancel the colour quenching effect by atest compound. The results are determined as percent inhibition ofenzyme activity. DMSO and 0.5 M EDTA are used as a control of 0% and100% inhibition, respectively. IC50 values are determined by non-linearcurve fit analysis using the OriginPro 6.1 program (OriginLab). In thisassay the compounds of formula I inhibit FAK activity at a IC₅₀<1 μM.Examples 188, 208 and 213 show IC₅₀ values of 15 nM, 1 nM and 7 nMrespectively.

The Biot-Y397 peptide (Biotin-SETDDYAEIID ammonium salt, SEQ ID NO:2) isdesigned to have the same amino acid sequence as the region from S392 toD402 of human (GenBank Accession Number L13616) and is prepared bystandard methods.

Purified recombinant hexahistidine-tagged human FAK kinase domain isobtained in the following way: Full-length human FAK cDNA is isolated byPCR amplification from human placenta Marathon-Ready™ cDNA (Clontech,No. 7411-1) with the 5′ PCR primer (ATGGCAGCTGCTTACCTTGAC, SEQ ID NO:3)and the 3′ PCR primer (TCAGTGTGGTCTCGTCTGCCC, SEQ ID NO:4) and subclonedinto a pGEM-T vector (Promega, No. A3600). After digestion with AccIII,the purified DNA fragment is treated with Klenow fragment. The cDNAfragment is digested with BamHI and cloned into pFastBacHTb plasmid(Invitrogen Japan K.K., Tokyo) previously cut with BamHI and Stu I. Theresultant plasmid, hFAK KD (M384-G706)/pFastBacHTb, is sequenced toconfirm its structure. The resulting DNA encodes a 364 amino acidprotein containing a hexahistidine tag, a spacer region and a rTEVprotease cleavage site at the N-terminal and the kinase domain of FAK(Met384-Gly706) from position 29 to 351.

Donor plasmid is transposed into the baculovirus genome, usingMaxEfficacy DH10Bac E. coli cells. Bacmid DNA is prepared by a simplealkaline lysis protocol described in the Bac-to-Bac® BaculovirusExpression system (Invitrogen). Sf9 insect cells are transfected basedon the protocol provided by the vendor (CeIIFECTIN®, Invitrogen). Theexpression of FAK in each lysate is analysed by SDS-PAGE and Westernblotting with anti-human FAK monoclonal antibody (clone #77 fromTransduction Laboratories).

The virus clone that shows the highest expression is further amplifiedby infection to Sf9 cells. Expression in ExpresSF+® cells (ProteinSciences Corp., Meriden, Conn., USA) gives high level of protein withlittle degradation. Cell lysates are loaded onto a column of HiTrap™Chelating Sepharose HP (Amersham Biosciences) charged with nickelsulfate and equilibrated with 50 mM HEPES pH 7.5, 0.5 M NaCl and 10 mMimidazole. Captured protein is eluted with increasing amounts ofimidazole in HEPES buffer/NaCl, and further purified by dialysis in 50mM HEPES pH 7.5, 10% glycerol and 1 mM DTT.

4. Phosphorylation Levels of FAK

Phosphorylation levels of FAK at Tyr397 are quantified by the sandwichELISA. Mouse mammary carcinoma 4T1 cells (1×10⁵) are plated in wells of96-well culture plates and incubated with or without variousconcentrations of a compound of formula I for 1 h in Dulbecco's modifiedeagle medium containing 10% FBS. The medium is removed and cells arelysed in 200 μL 50 mM Tris-HCl, pH 7.4, containing 1% NP-40, 0.25%sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 1 mM Na₃VO₄, 1mM NaF, 1 μg/mL aprotinin, 1 μg/mL leupeptin and 1 μg/mL pepstatin.After centrifugation, the supernatants are subjected to a sandwich ELISAto quantify the phosphorylated FAK and total FAK. Cell lysates areapplied to 96-well flat-bottom ELISA plates which have been pre-coatedwith 100 μL/well of 4 μg/mL mouse monoclonal anti-FAK antibody (clone77, Becton Dickinson Transduction Laboratories) in 50 mM Tris-HCl, pH9.5, containing 150 mM NaCl for 18 h at 4° C. and blocked with 300 μL ofBlockAce (Dainippon Pharmaceuticals Co.) diluted at 1:4 with H₂O at roomtemperature for 2 h. After washing with TBSN (20 mM Tris-HCl, pH 8.3,containing 300 mM NaCl, 0.1% SDS and 0.05% NP-40), total FAK is detectedwith 100 μL of 1 μg/ml anti-FAK polyclonal antibody (#65-6140, UpstateBiology Inc.), and phosphorylated FAK is detected with 100 μL of 0.25μg/μL anti-phosphorylated FAK (Y397) antibody (Affinity BioReagents,#OPA1-03071) in BlockAce diluted at 1:10 with H₂O. After 1 h incubationat room temperature, plates are washed with TBSN and 100 μL ofbiotinylated anti-rabbit IgG (#65-6140, Zymed Laboratolies Inc.) dilutedat 1:2000 with BlockAce diluted at 1:10 with H₂O is incubated at roomtemperature for 1 h. After washing with TBSN, ABTS solution substratekit (#00-2011, Zymed Lobolatories Inc.) is used for color development.Absorbance at 405 nm is measured after 20 min incubation at roomtemperature. The concentration of compound causing 50% reduction ofphosphorylation level of FAK (IC₅₀) is determined. In this assay,compounds of formula I reduce phosphorylation at an IC₅₀ of <1 μM.Examples 190, 198 and 210 show IC₅₀ values of 0.44 μM, 0.043 μM and 0.01μM respectively.

5. Anchorage-Independent Tumor Cell Growth Assay

Mouse mammary carcinoma 4T1 cells (5×10³) are plated in 96-well Ultralow Attachment plates (#3474, Corning Inc.) in 100 μL of Dulbecco'smodified eagle medium containing 10% FBS. Cells are cultured for 2 h andinhibitors are added at various concentrations in a final concentrationof 0.1% DMSO. After 48 h, cell growth is assayed with the cell countingkit-8 (Wako Pure Chemical), which uses a water soluble tetrazolium saltWST8. Twenty μL of the reagent is added into each well and cells arefurther cultured for 2 h. The optical density is measured at 450 nm. Theconcentration of compound causing 50% inhibition of growth may thus bedetermined. Examples 204, 213 and 206 show IC₅₀ values of 0.4 μM, 0.016μM and 0.09 μM respectively.

The compounds of the invention are therefore useful in the prevention ortreatment of disorders or diseases where ZAP-70 inhibition, and/or Sykinhibition play a role, e.g. diseases or disorders mediated by Tlymphocytes, B lymphocytes, mast cells and/or eosinophils e.g. acute orchronic rejection of organ or tissue allo- or xenografts,atheriosclerosis, vascular occlusion due to vacular injury such asangioplasty, restenosis, hypertension, heart failure, chronicobstructive pulmonary disease, CNS disease such as Alzheimer disease oramyotrophic lateral sclerosis, cancer, infectious disease such as AIDS,septic shock or adult respiratory distress syndrome,ischemia/reperfusion injury e.g. myocardial infarction, stroke, gutischemia, renal ailure or hermorrhage shock, or traumatic shock. Theagent of the invention are also useful in the treatment and/orprevention of acute or chronic inflammatory diseases or disorders orautoimmune diseases e.g. rheumatoid arthritis, osteoarthritis, systemiclupus erythematosus, Hashimoto's thyroidis, multiple sclerosis,myasthenia gravis, diabetes (type I and II) and the disorders associatedwith therewith, respiratory diseases such as asthma or inflammatoryliver injury, inflammatory glomerular injury, cutaneous manifestationsof immunologically-mediated disorders or illnesses, inflammatory andhyperproliferative skin diseases (such as psoriasis, atopic dermatitis,allergic contact dermatitis, irritant contact dermatitis and furthereczematous dermatitises, seborrhoeic dermatitis), inflammatory eyediseases, e.g. Sjoegren's syndrome, keratoconjunctivitis or uveitis,inflammatory bowel disease, Crohn's disease or ulcerative colitis.

Compounds of the invention are also useful in the prevention ortreatment of conditions caused by a malfunction of signal cascadesconnected with FAK, e.g. tumors, for example brain and other centralnervous system tumors (eg. tumors of the meninges, brain, spinal cord,cranial nerves and other parts of central nervous system, e.g.glioblastomas or medulla blastomas); head and/or neck cancer; breasttumors; circulatory system tumors (e.g. heart, mediastinum and pleura,and other intrathoracic organs, vascular tumors and tumor-associatedvascular tissue); excretory system tumors (e.g. kidney, renal pelvis,ureter, bladder, other and unspecified urinary organs); gastrointestinaltract tumors (e.g. oesophagus, stomach, small intestine, colon,colorectal, rectosigmoid junction, rectum, anus and anal canal), tumorsinvolving the liver and intrahepatic bile ducts, gall bladder, other andunspecified parts of biliary tract, pancreas, other and digestiveorgans); head and neck; oral cavity (lip, tongue, gum, floor of mouth,palate, and other parts of mouth, parotid gland, and other parts of thesalivary glands, tonsil, oropharynx, nasopharynx, pyriform sinus,hypopharynx, and other sites in the lip, oral cavity and pharynx);reproductive system tumors (e.g. vulva, vagina, Cervix uteri, Corpusuteri, uterus, ovary, and other sites associated with female genitalorgans, placenta, penis, prostate, testis, and other sites associatedwith male genital organs); respiratory tract tumors (e.g. nasal cavityand middle ear, accessory sinuses, larynx, trachea, bronchus and lung,e.g. small cell lung cancer or non-small cell lung cancer); skeletalsystem tumors (e.g. bone and articular cartilage of limbs, bonearticular cartilage and other sites); skin tumors (e.g. malignantmelanoma of the skin, non-melanoma skin cancer, basal cell carcinoma ofskin, squamous cell carcinoma of skin, mesothelioma, Kaposi's sarcoma);and tumors involving other tissues incluing peripheral nerves andautonomic nervous system, connective and soft tissue, retroperitoneumand peritoneum, eye and adnexa, thyroid, adrenal gland and otherendocrine glands and related structures, secondary and unspecifiedmalignant neoplasm of lymph nodes, secondary malignant neoplasm ofrespiratory and digestive systems and secondary malignant neoplasm ofother sites, tumors of blood and lymphatic system (e.g. Hodgkin'sdisease, Non-Hodgkin's lymphoma, Burkitt's lymphoma, AIDS-relatedlymphomas, malignant immunoproliferative diseases, multiple myeloma andmalignant plasma cell neoplasms, lymphoid leukemia, acute or chronicmyeloid leukemia, acute or chronic lymphocytic leukemia, monocyticleukemia, other leukemias of specified cell type, leukemia ofunspecified cell type, other and unspecified malignant neoplasms oflymphoid, haematopoietic and related tissues, for example diffuse largecell lymphoma, T-cell lymphoma or cutaneous T-cell lymphoma). Myeloidcancer includes e.g. acute or chronic myeloid leukaemia.

Where hereinbefore and subsequently a tumor, a tumor disease, acarcinoma or a cancer is mentioned, also metastasis in the originalorgan or tissue and/or in any other location are implied alternativelyor in addition, whatever the location of the tumor and/or metastasis is.

The compositions of the invention may be administered by anyconventional route, in particular parenterally, for example in the formof injectable solutions or suspensions, enterally, e.g. orally, forexample in the form of tablets or capsules, topically, e.g. in the formof lotions, gels, ointments or creams, or in a nasal or a suppositoryform. Pharmaceutical compositions comprising an agent of the inventionin association with at least one pharmaceutical acceptable carrier ordiluent may be manufactured in conventional manner by mixing with apharmaceutically acceptable carrier or diluent. Unit dosage forms fororal administration contain, for example, from about 0.1 mg to about 500mg of active substance. Topical administration is e.g. to the skin. Afurther form of topical administration is to the eye.

The compounds of formula I may be administered in free form or inpharmaceutically acceptable salt form, e.g. as indicated above. Suchsalts may be prepared in conventional manner and exhibit the same orderof activity as the free compounds.

In accordance with the foregoing, the present invention also provides:

(1) A compound of formula I or a pharmaceutically acceptable saltthereof, for use as a pharmaceutical;(2) A compound of formula I or a pharmaceutically acceptable saltthereof, for use as a ZAP-70, FAK and/or Syk tyrosine kinase inhibitor,for example for use in any of the particular indications hereinbeforeset forth;(3) A pharmaceutical composition, e.g. for use in any of the indicationsherein before set forth, comprising a compound of formula I or apharmaceutically acceptable salt thereof, together with one or morepharmaceutically acceptable diluents or carriers therefor.(4) A method for the treatment of any of particular indicationhereinbefore set forth in a subject in need thereof which comprisesadministering an effective amount of a compound of formula I or apharmaceutically acceptable salt thereof;(5) The use of a compound of formula I or a pharmaceutically acceptablesalt thereof, for the manufacture of a medicament for the treatment orprevention of a disease or condition in which ZAP-70, FAK and/or Syktyrosine kinase activation plays a role or is implicated; e.g. asdiscussed above.

Compounds of the invention may be administered as the sole activeingredient or together with other drugs useful against neoplasticdiseases, inflammatory disorders or in immunomodulating regimens. Forexample, the compounds of the invention may be used in combination withan active agent effective in various diseases as described above, e.g.with cyclosporins, rapamycins or ascomycins, or their immunosuppressiveanalogs or derivatives, e.g. cyclosporin A, cyclosporin G, Is a tx247,FK-506, sirolimus or everolimus; CCI-779, ABT578, AP23573,corticosteroids e.g. prednisone; cyclophosphamide; azathioprene;methotrexate; gold salts, sulfasalazine, antimalarials; leflunomide;mizoribine; mycophenolic acid; mycophenolate mofetil;15-deoxyspergualine; an EDG receptor agonist having acceleratinglymphocyte homing activity, e.g FTY720 or an analogue thereof,immuno-suppressive monoclonal antibodies, e.g. monoclonal antibodies toleukocyte receptors, e.g. MHC, CD2, CD3, CD4, CD7, CD25, CD28, CD40,CD45, CD58, CD80, CD86, CD152, CD137, CD154, ICOS, LFA-1, VLA-4 or theirligands; or other immunomodulatory compounds, e.g. CTLA4Ig.

A compound of formula I may also be used to advantage in combinationwith other antiproliferative agents. Such antiproliferative agentsinclude, but are not limited to aromatase inhibitors, antiestrogens,topoisomerase I inhibitors, topoisomerase II inhibitors, microtubuleactive agents, alkylating agents, histone deacetylase inhibitors,farnesyl transferase inhibitors, COX-2 inhibitors, MMP inhibitors, mTORinhibitors, antineoplastic antimetabolites, platin compounds, compoundsdecreasing the protein kinase activity and further anti-angiogeniccompounds, gonadorelin agonists, anti-androgens, bengamides,bisphosphonates, antiproliferative antibodies and temozolomide(TEMODAL®).

The term “aromatase inhibitors” as used herein relates to compoundswhich inhibit the estrogen production, i.e. the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to steroids,especially exemestane and formestane and, in particular, non-steroids,especially aminoglutethimide, vorozole, fadrozole, anastrozole and, veryespecially, letrozole. A combination of the invention comprising anantineoplastic agent which is an aromatase inhibitor may particularly beuseful for the treatment of hormone receptor positive breast tumors.

The term “antiestrogens” as used herein relates to compounds whichantagonize the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride.

The term “topoisomerase I inhibitors” as used herein includes, but isnot limited to topotecan, irinotecan, 9-nitrocamptothecin and themacromolecular camptothecin conjugate PNU-166148 (compound A1 inWO99/17804).

The term “topoisomerase II inhibitors” as used herein includes, but isnot limited to the antracyclines doxorubicin (including liposomalformulation, e.g. CAELYX™), epirubicin, idarubicin and nemorubicin, theanthraquinones mitoxantrone and losoxantrone, and the podophillotoxinesetoposide and teniposide.

The term “microtubule active agents” relates to microtubule stabilizingand microtubule destabilizing agents including, but not limited to thetaxanes paclitaxel and docetaxel, the vinca alkaloids, e.g.,vinblastine, especially vinblastine sulfate, vincristine especiallyvincristine sulfate, and vinorelbine, discodermolide and epothilones,such as epothilone B and D.

The term “alkylating agents” as used herein includes, but is not limitedto cyclophosphamide, ifosfamide and melphalan.

The term “histone deacetylase inhibitors” relates to compounds whichinhibit the histone deacetylase and which possess antiproliferativeactivity.

The term “farnesyl transferase inhibitors” relates to compounds whichinhibit the farnesyl transferase and which possess antiproliferativeactivity.

The term “COX-2 inhibitors” relates to compounds which inhibit thecyclooxygenase type 2 enyzme (COX-2) and which possess antiproliferativeactivity such as celecoxib (Celebrex®), rofecoxib (Vioxx®) andlumiracoxib (COX189).

The term “MMP inhibitors” relates to compounds which inhibit the matrixmetalloproteinase (MMP) and which possess antiproliferative activity.

The term “antineoplastic antimetabolites” includes, but is not limitedto 5-fluorouracil, tegafur, capecitabine, cladribine, cytarabine,fludarabine phosphate, fluorouridine, gemcitabine, 6-mercaptopurine,hydroxyurea, methotrexate, edatrexate and salts of such compounds, andfurthermore ZD 1694 (RALTITREXED™), LY231514 (ALIMTA™), LY264618(LOMOTREXOL™) and OGT719.

The term “platin compounds” as used herein includes, but is not limitedto carboplatin, cis-platin and oxaliplatin.

The term “compounds decreasing the protein kinase activity and furtheranti-angiogenic compounds” as used herein includes, but is not limitedto compounds which decrease the activity of e.g. the VascularEndothelial Growth Factor (VEGF), the Epidermal Growth Factor (EGF),c-Src, protein kinase C, Platelet-derived Growth Factor (PDGF), Bcr-Abltyrosine kinase, c-kit, Flt-3 and Insulin-like Growth Factor I Receptor(IGF-IR) and Cyclin-dependent kinases (CDKs), and anti-angiogeniccompounds having another mechanism of action than decreasing the proteinkinase activity.

Compounds which decrease the activity of VEGF are especially compoundswhich inhibit the VEGF receptor, especially the tyrosine kinase activityof the VEGF receptor, and compounds binding to VEGF, and are inparticular those compounds, proteins and monoclonal antibodiesgenerically and specifically disclosed in WO 98/35958 (describingcompounds of formula I), WO 00/09495, WO 00/27820, WO 00/59509, WO98/11223, WO 00/27819, WO 01/55114, WO 01/58899 and EP 0 769 947; thoseas described by M. Prewett et al in Cancer Research 59 (1999) 5209-5218,by F. Yuan et al in Proc. Natl. Acad. Sci. USA, vol. 93, pp.14765-14770, December 1996, by Z. Zhu et al in Cancer Res. 58, 1998,3209-3214, and by J. Mordenti et al in Toxicologic Pathology, vol. 27,no. 1, pp 14-21, 1999; in WO 00/37502 and WO 94/10202; Angiostatin™,described by M. S. O'Reilly et al, Cell 79, 1994, 315-328; andEndostatin™, described by M. S. O'Reilly et al, Cell 88, 1997, 277-285;

compounds which decrease the activity of EGF are especially compoundswhich inhibit the EGF receptor, especially the tyrosine kinase activityof the EGF receptor, and compounds binding to EGF, and are in particularthose compounds generically and specifically disclosed in WO 97/02266(describing compounds of formula IV), EP 0 564 409, WO 99/03854, EP0520722, EP 0 566 226, EP 0 787 722, EP 0 837 063, WO 98/10767, WO97/30034, WO 97/49688, WO 97/38983 and, especially, WO 96/33980;compounds which decrease the activity of c-Src include, but are notlimited to, compounds inhibiting the c-Src protein tyrosine kinaseactivity as defined below and to SH2 interaction inhibitors such asthose disclosed in WO97/07131 and WO97/08193;compounds inhibiting the c-Src protein tyrosine kinase activity include,but are not limited to, compounds belonging to the structure classes ofpyrrolopyrimidines, especially pyrrolo[2,3-d]pyrimidines, purines,pyrazopyrimidines, especially pyrazo[3,4-d]pyrimidines,pyrazopyrimidines, especially pyrazo[3,4-d]pyrimidines andpyridopyrimidines, especially pyrido[2,3-d]pyrimidines. Preferably, theterm relates to those compounds disclosed in WO 96/10028, WO 97/28161,WO97/32879 and WO97/49706;compounds which decreases the activity of the protein kinase C areespecially those staurosporine derivatives disclosed in EP 0 296 110(pharmaceutical preparation described in WO 00/48571) which compoundsare protein kinase C inhibitors;further specific compounds that decrease protein kinase activity andwhich may also be used in combination with the compounds of the presentinvention are Imatinib (Gleevec®/Glivec®), PKC412, Iressa™ (ZD1839),PK1166, PTK787, ZD6474, GW2016, CHIR-200131, CEP-7055/CEP-5214,CP-547632 and KRN-633;anti-angiogenic compounds having another mechanism of action thandecreasing the protein kinase activity include, but are not limited toe.g. thalidomide (THALOMID), celecoxib (Celebrex), SU5416 and ZD6126.

The term “gonadorelin agonist” as used herein includes, but is notlimited to abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274.

The term “anti-androgens” as used herein includes, but is not limited tobicalutamide (CASODEX™), which can be formulated, e.g. as disclosed inU.S. Pat. No. 4,636,505.

The term “bengamides” relates to bengamides and derivatives thereofhaving aniproliferative properties.

The term “bisphosphonates” as used herein includes, but is not limitedto etridonic acid, clodronic acid, tiludronic acid, pamidronic acid,alendronic acid, ibandronic acid, risedronic acid and zoledronic acid.

The term “antiproliferative antibodies” as used herein includes, but isnot limited to trastuzumab (Herceptin™), Trastuzumab-DM1, erlotinib(Tarceva™), bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553(anti-CD40) and 2C4 Antibody.

The structure of the active agents identified by code nos., generic ortrade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g. PatentsInternational (e.g. IMS World Publications).

In accordance with the foregoing the present invention provides in a yetfurther aspect:

(6) A method as defined above comprising co-administration, e.g.concomitantly or in sequence, of a therapeutically effective amount ofa) a compound of formula I or a pharmaceutically acceptable saltthereof, and b) a second drug substance, said second drug substancebeing for example for use in any of the particular indicationshereinbefore set forth.(7) A combination comprising a therapeutically effective amount of aZAP-70, FAK and/or Syk tyrosine kinase inhibitor, e.g. a compound offormula I or a pharmaceutically acceptable salt thereof, and a seconddrug substance, said second drug substance being for example asdisclosed above.

Where a ZAP-70, FAK and/or Syk tyrosine kinase inhibitor, e.g. acompound of formula I, is administered in conjunction with otherimmunosuppressive/immunomodulatory, anti-inflammatory or antineoplasticagent, e.g. as disclosed above, dosages of the co-administered drug oragent will of course vary depending on the type of co-drug or -agentemployed, or the specific drug or agent used, or the condition beingtreated and so forth.

Representative FAK inhibitors are the compounds of Examples Nos. 187-203and 209-212.

1-10. (canceled)
 11. A compound of formula I

wherein X is ═CR⁰—; R⁰ is hydrogen; R² is —SO₂N(R¹⁰)R¹¹; each of R¹ andR³ independently is hydrogen; hydroxy; C₁-C₈alkyl; C₂-C₈alkenyl;C₃-C₈cycloalkyl; C₃-C₈cycloalkyl-C₁-C₈alkyl; hydroxyC₁-C₈alkyl;C₁-C₈alkoxyC₁-C₈alkyl; hydroxyC₁-C₈alkoxyC₁-C₈alkyl; arylC₁-C₈alkylwhich optionally may be substituted on the ring by hydroxy, C₁-C₈alkoxy,carboxy or C₁-C₈alkoxycarbonyl; or each of R¹ and R³, independently, ishalogen; halo-C₁-C₈alkyl; C₁-C₈alkoxy; halo-C₁-C₈alkoxy;hydroxyC₁-C₈alkoxy; C₁-C₈alkoxyC₁-C₈alkoxy; aryl; arylC₁-C₈alkoxy;heteroaryl; heteroaryl-C₁-C₄alkyl; 5 to 10 membered heterocyclic ring;nitro; carboxy; C₂-C₈alkoxycarbonyl; C₂-C₈alkylcarbonyl;—N(C₁-C₈alkyl)C(O)C₁-C₈alkyl; —N(R¹⁰)R¹¹; —CON(R¹⁰)R¹¹;or)-C₁-C₄-alkylene-SO₂N(R¹⁰)R¹¹; wherein each of R¹⁰ and R¹¹independently is hydrogen; hydroxy; C₁-C₈alkyl; C₂-C₈alkenyl;C₃-C₈cycloalkyl; C₃-C₈cycloalkyl-C₁-C₈alkyl; C₁-C₈alkoxyC₁-C₈alkyl;hydroxyC₁-C₈alkoxyC₁-C₈alkyl; hydroxyC₁-C₈alkyl; (C₁-C₈alkyl)-carbonyl;arylC₁-C₈alkyl which optionally may be substituted on the ring byhydroxy, C₁-C₈alkoxy, carboxy or C₂-C₈alkoxycarbonyl; or 5 to 10membered heterocyclic ring; R⁴ is hydrogen; R⁵ is hydrogen; halogen;C₁₋₄alkyl; or CF3; R⁶ is hydrogen; each of R⁷, R⁸ and R⁹ isindependently hydrogen; hydroxy; C₁-C₈alkyl; C₂-C₈alkenyl;halo-C₁-C₈alkyl; C₁-C₈alkoxy; C₃-C₈cycloalkyl;C₃-C₈cycloalkylC₁-C₈alkyl; arylC₁-C₈alkyl; —Y—R¹² wherein Y is a directbond or O and R¹² is a substituted or unsubstituted 5, 6 or 7 memberedheterocyclic ring comprising 1, 2 or 3 heteroatoms selected from N, Oand S; carboxy; (C₁-C₈alkoxy)-carbonyl; —N(C₁₋₈alkyl)-CO—NR¹⁰R¹¹;—CONR¹⁰R¹¹; —N(R¹⁰)(R¹¹); —SO₂N(R¹⁰)R¹¹; R⁷ and R⁸ or R⁸ and R⁹,respectively form together with the carbon atoms to which they areattached, a 5 or 6 membered heteroaryl comprising 1, 2 or 3 heteroatomsselected from N, O and S; or a 5 or 6 membered carbocyclic ring; whereinany alkyl, alkoxy, alkenyl, cycloalkyl, heterocyclic ring, aryl orheteroaryl may be unsubstituted or substituted by one or moresubstituents selected from halogen; OH; C₁-C₈alkyl; C₁-C₈alkoxy; nitro;cyano; COOH; carbamoyl; C(NH₂)═NOH; —N(R¹⁰)R¹¹; C₃-C₆cycloalkyl; 3 to 7membered heterocyclic ring; phenyl; phenyl-C₁₋₄alkyl; 5 or 6 memberedheteroaryl; in free form or salt form.
 12. A compound according to claim11 wherein at most one of R¹ or R³ is —CON(R¹⁰)R¹¹.
 13. A compoundaccording to claim 11 which is a compound of formula X₄

wherein R², R⁵, R⁷, R⁸ and R⁹ are as defined in claim
 11. 14. A processfor the production of a compound of formula I according to claim 11,comprising the steps of reacting a compound of formula II

wherein R¹, R², R³, R⁴, R⁵, R⁶ and X are as defined in claim 11, and Yis a leaving group; with a compound of formula III

wherein R⁷, R⁸ and R⁹ are as defined in claim 11; and recovering theresulting compound of formula I in free form or in salt form, and, whererequired, converting the compound of formula I obtained in free forminto the desired salt form, or vice versa.
 15. A compound according toclaim 11 in free form or in pharmaceutically acceptable salt form, foruse as a pharmaceutical.
 16. A pharmaceutical composition comprising acompound of formula I according to claim 11 or a pharmaceuticallyacceptable salt thereof, together with one or more pharmaceuticallyacceptable carriers or diluents therefor.
 17. A method for the treatmentor prevention of a disease or condition in which ZAP-70, FAK and/or Syktyrosine kinase activation plays a role or is implicated, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula I according to claim 11 in free form orin pharmaceutically acceptable salt form.
 18. A combination whichcomprises (a) a therapeutically effective amount of a compound offormula I according to claim 11 as a ZAP-70, FAK and/or Syk inhibitorand (b) a second drug substance.
 19. A combination which comprises (a) atherapeutically effective amount of a compound of formula I according toclaim 12 as a ZAP-70, FAK and/or Syk inhibitor and (b) a second drugsubstance.
 20. A combination which comprises (a) a therapeuticallyeffective amount of a compound of formula I according to claim 13 as aZAP-70, FAK and/or Syk inhibitor and (b) a second drug substance.